scholarly journals Design of a multiplex PCR method for detection of toxigenic-pathogenic in Vibrio cholerae

2013 ◽  
Vol 6 (2) ◽  
pp. 115-118 ◽  
Author(s):  
Fooladi AA Imani ◽  
Islamieh D Iman ◽  
Doust R Hosseini ◽  
A Karami ◽  
SM Marashi
Keyword(s):  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
Pcr Method

scholarly journals Rapid Method for Species-Specific Identification and Determination of Toxigenicity of Vibrio Cholerae from Natural Aquatic Environment

1970 ◽  
Vol 1 (1) ◽  
pp. 69-75
Author(s):  
Bidhan Chakraborty ◽  
Khadiza Zaman ◽  
Md Majibur Rahman
Keyword(s):  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
Serological Test ◽  
Seasonal Pattern ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
Pcr Multiplex ◽  
Pcr Method ◽  
Species Specific

Cholera caused by toxigenic Vibrio cholerae is a major public health problem confronting developing countries, where outbreaks occur in a regular seasonal pattern and are particularly associated with poverty and poor sanitation. It is generally accepted that seven distinct pandemics of cholera have occurred since the onset of the first pandemic in 1817. Again Vibrio cholerae is capable of surviving in aquatic environments for extended periods and is considered as autochthonous species in estuarine and brackish waters. Therefore, the present study was designed to isolate V. cholerae from natural environmental samples subsequently identified by conventional and molecular biological techniques. A total number of 10 isolates were included randomly in this study based on their initial identification. The serotypes of the isolates were determined by serological test (slide agglutination) and the number of serotypes O1, O139 and non-O1/O139 were 3, 2 and 5 respectively which were reconfirmed by PCR method. Finally, the toxigenicity of the isolates was analyzed by multiplex PCR method and five (5) isolates were found to contain the ctx gene, the major virulence factor of V. cholerae. Key Words: Vibrio cholerae, Simplex PCR, Multiplex PCR, Serotypes, Toxigenicity.   doi:10.3329/sjps.v1i1.1811 S. J. Pharm. Sci. 1(1&2): 69-75

scholarly journals Evaluation of new primers for detecting toxigenic vibrio cholerae by multiplex PCR

2011 ◽  
Vol 2 (1) ◽  
pp. 1
Author(s):  
Jalil F. Mehrabadi ◽  
Parisa Morsali ◽  
Hamideh Rohani nejad ◽  
Abbas Ali Imani Fooladi ◽  
Sarah Najafy
Keyword(s):  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
Salmonella Typhi ◽  
Dilution Method ◽  
Microbial Culture ◽  
E Coli ◽  
Multiplex Pcr Assay ◽  
Colony Forming Units ◽  
Sensitive Tool ◽  
Pcr Method

<em>Vibrio cholerae</em> is the etiological agent of cholera that has emerged as an endemic disease in different regions of the world in recent years. Traditional microbial culture and microscopy methods are considered to be the best standard for diagnosing <em>V. cholerae</em> infection. These methods, however, delay any available confirmatory answer by days. Molecular methods have the potential to provide sensitive, accurate, and rapid analysis of<em> V. cholerae</em> infection. We have developed a multiplex PCR assay to detect virulence and toxigenic-associated (VTA) genes (<em>ctxA, tcpA, and ompW</em>). To evaluate PCR specificity, additional bacteria from the enterobacteriaceae family (<em>Salmonella typhi, Shigella dysantry, and entrotoxigenic E. coli</em>) and <em>Aeromonas hidrophyla</em> were examined in this study. Specificity tests were evaluated using the genome dilution method. Importantly, the results show that our PCR specificity method represents the best tool for the rapid detection of VTA genes because of its simplicity, cost effectiveness, and accuracy. This multiplex PCR method can be used for examining the existence of VTA genes in patient samples, and therefore will distinguish <em>V. cholerae</em> from other vibrios and bacteria. This method is able to detect 10-100 colony forming units (CFUs) of V.Cholerae and 8.5-85 picograms (pg) of genomic DNA. The multiplex PCR method is also more specific and sensitive than other methods, validating it as an appropriate and sensitive tool for detecting the presence of toxigenic and pathogenic <em>V. cholerae</em>.

A novel common primer multiplex PCR (CP-M-PCR) method for the simultaneous detection of meat species

Food Control ◽  
2009 ◽  
Vol 20 (4) ◽  
pp. 366-370 ◽  
Author(s):  
Weibin Bai ◽  
Wentao Xu ◽  
Kunlun Huang ◽  
Yanfang Yuan ◽  
Sishuo Cao ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Meat Species ◽  
Pcr Method

Prevalence and Molecular Characterization of Vibrio cholerae from Ice and Beverages Sold in Jakarta, Indonesia, Using Most Probable Number and Multiplex PCR

2012 ◽  
Vol 75 (4) ◽  
pp. 651-659 ◽  
Author(s):  
DIANA E. WATURANGI ◽  
NATANIA PRADITA ◽  
JESSICA LINARTA ◽  
SWAPAN BANERJEE
Keyword(s):  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
Virulence Gene ◽  
Watery Diarrhea ◽  
Most Probable Number ◽  
Intestinal Infection ◽  
Safety Measures ◽  
Probable Number ◽  
Tropical Regions

Vibrio cholerae is well recognized as the causative agent of cholera, an acute intestinal infection characterized by watery diarrhea that may lead to dehydration and death in some cases. V. cholerae is a natural inhabitant of the aquatic environment in the tropical regions. Jakarta has the highest percentage of individuals affected by sporadic diarrheal illness compared with other areas in Indonesia. Inadequate safety measures for drinking water supplies, improper sanitation, and poor hygiene can increase the risk of cholera outbreaks. Few studies have been conducted on the prevalence of these bacteria in ice and beverages that are popularly sold and consumed in Jakarta. In this study, we detected and quantified V. cholerae from ice and beverages collected from several areas in five regions of Jakarta. Levels of V. cholerae in both ice and beverages were determined with the three-tube most-probable-number (MPN) method and ranged from &lt;0.3 to &gt;110 MPN/ml. The presence of regulatory and virulence gene sequences was determined by using uniplex and multiplex PCR assays. Of 110 samples tested, 33 (30%) were positive for V. cholerae; 21 (64%) were ice samples and the remaining 12 (36%) were beverages. A total of 88 V. cholerae strains were isolated, based on the presence of the toxR gene sequence identified by PCR. Other genetic markers, such as hlyA (59%), ompU (16%), and ctxA (19%), also were found during the search for potential pathogenic strains. The detection and isolation of potentially harmful V. cholerae from ice and beverages in Jakarta indicate that these products pose a health risk from choleragenic vibrios, particularly because of the emergence of classical biotypes of V. cholerae O1 and potentially harmful non-O1 serovars of this species.

scholarly journals Modified Multiplex PCR Method for Detection of Pyrogenic Exotoxin Genes in Staphylococcal Isolates

2004 ◽  
Vol 42 (8) ◽  
pp. 3869-3872 ◽  
Author(s):  
A. Lovseth ◽  
S. Loncarevic ◽  
K. G. Berdal
Keyword(s):  
Multiplex Pcr ◽  
Pcr Method

Prevalence, Antibiogram, and cdt Genes of Toxigenic Campylobacter jejuni in Salad Style Vegetables (Ulam) at Farms and Retail Outlets in Terengganu

2015 ◽  
Vol 78 (1) ◽  
pp. 65-71 ◽  
Author(s):  
MOHD IKHSAN KHALID ◽  
JOHN YEW HUAT TANG ◽  
NABILA HUDA BAHARUDDIN ◽  
NASIHA SHAKINA RAHMAN ◽  
NURUL FAIZZAH RAHIMI ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Multiplex Pcr ◽  
Campylobacter Jejuni ◽  
Antibiotic Susceptibility ◽  
Penicillin G ◽  
Diffusion Method ◽  
Cytolethal Distending Toxin ◽  
Disk Diffusion Method ◽  
Retail Outlets ◽  
Pcr Method

The present study was conducted to investigate the prevalence and antibiotic resistance among Campylobacter jejuni in ulam at farms and retail outlets located in Kuala Terengganu, Malaysia. A total of 526 samples (ulam, soil, and fertilizer) were investigated for the presence of C. jejuni and the gene for cytolethal distending toxin (cdt) by using a multiplex PCR method. Antibiotic susceptibility to 10 types of antibiotics was determined using the disk diffusion method for 33 C. jejuni isolates. The average prevalence of contaminated samples from farms, wet markets, and supermarkets was 35.29, 52.66, and 69.88%, respectively. The cdt gene was not detected in 24 of the 33 C. jejuni isolates, but 9 isolates harbored cdtC. Antibiotic resistance in C. jejuni isolates was highest to penicillin G (96.97% of isolates) followed by vancomycin (87.88%), ampicillin (75.76%), erythromycin (60.61%), tetracycline (9.09%), amikacin (6.06%), and norfloxacin (3.03%); none of the isolates were resistant to ciprofloxacin, enrofloxacin, and gentamicin. In this study, C. jejuni was present in ulam, and some isolates were highly resistant to some antibiotics but not to quinolones. Thus, appropriate attention and measures are required to prevent C. jejuni contamination on farms and at retail outlets.

Investigation of Children with Acute Gastroenteritis by Multiplex PCR Method in Central Part of Turkey

2022 ◽  
Author(s):  
Fatih Yılmaz ◽  
Havva Kaya ◽  
Mehmet Özdemir
Keyword(s):  
Multiplex Pcr ◽  
Acute Gastroenteritis ◽  
Hospital Admissions ◽  
Age Groups ◽  
Viral Gastroenteritis ◽  
Viral Agent ◽  
Mortality And Morbidity ◽  
Stool Samples ◽  
Pcr Method ◽  
Norovirus Gii

Abstract Objective Gastroenteritis is a disease that affects all age groups, especially children, and causes high mortality and morbidity in all countries. The most common agents of acute gastroenteritis are viral agents. As a result, millions of diarrhea attacks and hospital admissions occur worldwide every year due to viral gastroenteritis. This study uses the multiplex polymerase chain reaction (PCR) method to investigate the viruses that are the causative agents of viral gastroenteritis in the pediatric patient group in Konya, Turkey. Methods Stool samples of 94 patients aged 0 to 18 years sent from Emergency clinics and Pediatric outpatient clinics, Meram Medical Faculty Hospital Pediatric clinics, Konya Necmettin Erbakan University to Medical Microbiology Laboratory with a diagnosis of gastroenteritis between February and December 2018 were included in the study. Stool samples were stored at –80°C until the time of the analysis. Deoxyribonucleic acid/ribonucleic acid isolation from stool samples was performed with EZ1 Virus Mini Kit v2.0 (Qiagen, Hilden, Germany) using an automatic extraction system (BioRobot EZ1 system, Qiagen). The presence of astrovirus, rotavirus, adenovirus, norovirus (GI, GII), and sapovirus agents was investigated by the multiplex PCR method (Fast Track Diagnostics, Luxembourg) viral gastroenteritis kit. Results Viral gastroenteritis agents were detected in 56.3% of the patients. One viral agent was detected in 47 (50%) of these patients and at least two viral agents in 6 (6.3%) of them. Norovirus GII was detected in 20 (21.2%) of the children included in the study, adenovirus in 13 (13.8%), rotavirus in 11 (12.8%), astrovirus in 11 (11.7%), sapovirus in 4 (4.2%), and norovirus GI in 1 (1.06%). When the distribution of viral agents was examined by months, the most number of agents were observed (21; 35%) in May, followed by April and June (12; 20%). Considering the distribution of the prevalence of the agents by age, it was seen to be mainly between 0 and 12 months (42%). Conclusion Considering that the most common viral agent in our region is norovirus GII, it will be useful to investigate the norovirus that is not routinely examined in children who are admitted to clinics with the complaint of gastroenteritis. It will be appropriate to examine routinely adenovirus, rotavirus, and norovirus in the laboratory, especially in children with diarrhea and vomiting in the winter and spring months.

Mycobacterium tuberculosis typing using Allele-specific oligonucleotide multiplex PCR (ASO–PCR) method

2021 ◽  
Author(s):  
Maryam Shafipour ◽  
Hesamaddin Shirzad-Aski ◽  
Ezzat Allah Ghaemi ◽  
Ahmad Sohrabi ◽  
Maya Babaii Kochaksaraei ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Multiplex Pcr ◽  
Allele Specific ◽  
Pcr Method
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